title: Metagenomics for NoV surveillance: Outbreak-related sample sequencing
description: For Norovirus (NoV) genomic surveillance, metagenomics sequencing of viral nucleotides captured by VirCapSeq-VERT probe set, was performed. This metagenomics approach, in theory agnostic, offers the great advantage of unbiased detection of NoV, including the detection of novel NoV recombinants. Samples come from three different sequencing sets - Baseline Survey (BLS) samples, Outbreak-related (OB) samples, and two synthetic Test Set samples (TSS1 and TSS2). The samples from norovirus outbreaks associated with oysters included the clinical human stool samples from ill consumers and samples from the oyster batch linked to the outbreak were selected. The filtrate containing nucleic acids was treated with 10 µL/mL OmniCleave™ Endonuclease (Lucigen Corporation) and 100 µL MgCl2 (100mM) for 1h at 37°C. Nucleic acids extraction was performed using the NucliSens Extraction Reagents (bioMérieux, France) according to the manufacturer's instructions to obtain 100µL of nucleic acids in elution buffer. An additional RNA purification was performed using the RNA Clean & Concentrator TM-5 kit (Zymo Research, Irvine USA) (Strubbia et al., 2019) to remove traces of inhibitors and endonucleases.